Construction and bacteriostatic effect analyses of a recombinant thermostable Newcastle disease virus expressing cecropin AD

Lun Yao; Xiangfei Ren; Ping Zhou; Junjie Yang; Chi Zeng; Zhe Zeng; Yu Shang; Helong Feng; Mengyun Jin; Qianni Xiao; Huabin Shao; Qingping Luo; Sishun Hu; Guoyuan Wen

doi:10.1016/j.vetmic.2025.110397

Construction and bacteriostatic effect analyses of a recombinant thermostable Newcastle disease virus expressing cecropin AD

Vet Microbiol. 2025 Jan 17:302:110397. doi: 10.1016/j.vetmic.2025.110397. Online ahead of print.

Authors

Lun Yao¹, Xiangfei Ren², Ping Zhou³, Junjie Yang², Chi Zeng⁴, Zhe Zeng¹, Yu Shang¹, Helong Feng¹, Mengyun Jin¹, Qianni Xiao¹, Huabin Shao¹, Qingping Luo⁵, Sishun Hu⁶, Guoyuan Wen⁷

Affiliations

¹ Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China; Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture and Rural Affairs), Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China.
² Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430064, China; Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture and Rural Affairs), Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China.
³ Hubei Institute of Veterinary Durg Control, Wuhan 430064, China.
⁴ School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, China.
⁵ Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China; Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture and Rural Affairs), Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China; Hubei Hongshan Laboratory, Wuhan 430064, China.
⁶ College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430064, China. Electronic address: [email protected].
⁷ Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China; Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture and Rural Affairs), Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China. Electronic address: [email protected].

PMID: 39862796
DOI: 10.1016/j.vetmic.2025.110397

Abstract

Cecropin AD (CAD), a hybrid antimicrobial peptide composed of the first 11 residues of cecropin A and last 26 residues of cecropin D, is a promising antibiotic candidate. Therefore, an efficient and convenient method for producing CAD is necessary for commercial applications. The Newcastle disease virus (NDV) has been widely used as a platform for gene delivery and exogenous protein expression. In this study, we constructed a recombinant NDV that expresses CAD. To obtain high expression of the CAD peptide, tandem repeats of the cad gene were inserted into the genomes of the thermostable NDV strain TS09-C using reverse genetic technology. The thermostable recombinant NDV, namely rTS-CAD₃, showed thermostability and growth kinetics similar to those of their parental strain. A bacteriostatic test showed that rTS-CAD₃ inhibited Staphylococcus aureus (gram-positive bacteria) and Escherichia coli (gram-negative bacteria) in vitro. We further determined the bacteriostatic effects of rTS-CAD₃ expressed CAD against S. aureus in skin wound infections. The results showed that rTS-CAD₃ subcutaneously injection improved wound healing and reduced S. aureus decolonization. In summary, our results indicate that the rTS-CAD₃ expressing CAD peptide is a potent antimicrobial agent against S. aureus and E. coli and may be applied to accelerate wound healing in farm animals.

Keywords: Cecropin AD; Newcastle disease virus; Staphylococcus aureus; Wound healing.